ICAR-National Institute for Plant Biotechnology, New Delhi, India.
Department of Biotechnology, Kumaun University, Nainital, India.
Pest Manag Sci. 2022 Feb;78(2):803-811. doi: 10.1002/ps.6694. Epub 2021 Nov 12.
The nefarious hemipteran mustard aphid (Lipaphis erysimi) inflicts colossal yield losses in Brassica crops including Indian mustard (Brassica juncea). Lack of an accessible resistance source has been the primary impediment in breeding varietal resistance against aphids. In recent years, in planta RNAi-mediated resistance has been demonstrated in model plants as a potential tool for protection against insect pests. However, translational application in crop species is imperative for critical assessment of this technology in breeding effective resistance.
The essential role of sucrase 1 (SUC1) in mitigating osmotic pressure imposed by sucrose-rich phloem sap inside the insect gut is corroborated by its expression pattern in L. erysimi. Transgenic lines of Indian mustard were developed expressing SUC1 hairpin RNA for its host-mediated delivery into the infesting aphids. The expression of the dsRNA encoding cassette, and generation of siRNA molecules in transgenic B. juncea lines were verified by quantitative reverse transcription (RT)-PCR, stem-loop RT-PCR and Northern hybridization. Rearing of L. erysimi on the transgenic lines resulted in 22-40% reduction in aphid fecundity. The observed retardation in aphid reproduction was coherent with the detection of SUC1-specific siRNA molecules and attenuation of the SUC1 transcript level in L. erysimi fed on the transgenic lines.
Augmenting varietal resistance can substantially reduce usage of toxic agrochemicals in crop protection. This attempt was the first successful demonstration of host-mediated RNAi of an aphid gene in any Brassica crop. It paves the way for more rigorous attempt of engineering RNAi-based resistance against aphids in Brassica crops. © 2021 Society of Chemical Industry.
恶毒的半翅目芥菜蚜(Lipaphis erysimi)在包括芥菜(Brassica juncea)在内的 Brassica 作物上造成了巨大的产量损失。缺乏易于获得的抗性来源一直是培育抗蚜品种的主要障碍。近年来,在模式植物中已证明体内 RNAi 介导的抗性是一种潜在的工具,可以防止昆虫害虫侵害。然而,该技术在作物中的转化应用对于在育种中评估这种技术的有效性至关重要。
蔗糖酶 1(SUC1)在减轻昆虫肠道内富含蔗糖的韧皮部汁液所施加的渗透压方面起着重要作用,这一点得到了 L.erysimi 中其表达模式的证实。通过表达 SUC1 发夹 RNA 来开发芥菜的转基因品系,用于其在侵染蚜虫中的宿主介导传递。dsRNA 编码盒的表达和转基因 B. juncea 品系中的 siRNA 分子的产生通过定量反转录(RT)-PCR、茎环 RT-PCR 和 Northern 杂交进行了验证。在转基因品系上饲养 L.erysimi 导致蚜虫繁殖力降低 22-40%。在以转基因系为食的蚜虫中检测到 SUC1 特异性 siRNA 分子和 SUC1 转录本水平的衰减,这与观察到的蚜虫繁殖延迟一致。
增加品种抗性可以大大减少作物保护中有毒农药的使用。这项尝试是首例在任何 Brassica 作物中成功进行的蚜虫基因的宿主介导 RNAi 演示。它为在 Brassica 作物中基于 RNAi 的工程抗性来防治蚜虫奠定了基础。© 2021 化学工业协会。